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Fetal Bovine Serum (FBS) has been a vital and historically irreplaceable serum supplement for cell culture media. Limitations including inherent contaminants and batch inconsistency alongside high and volatile prices are well known to scientists. This makes its use undesirable in research and clinical settings.
Media City Scientific’s Fetal Bovine Serum Replacement Solution (FRS™ Pioneer) is a drop-in chemically defined and animal component free FBS replacement. FRS™ Pioneer supports the growth of a broad range of cell types. |
FRS™ Pioneer is a drop-in, chemically defined FBS replacement that is: > Cost effective – priced on par with premium FBS > Consistent – enabling scientific reproducibility and reliability > Animal-free – contains no animal derived components > Effective – supports a broad range of cells at a similar level of performance to FBS |
Application Notes and Recommendations
The following notes provide recommendations for use of FRS™ Pioneer and highlight key points of difference between the application of FRS™ Pioneer and FBS.
Recommendations for cell culture using FRS™ Pioneer
Optimising culture conditions:
- Thawing: Thaw FRS™ Pioneer in the waterbath. Swirl regularly and remove from heat immediately following thaw. Prepare aliquots and store at –20. Thawing in the fridge is not recommended.
- Antibiotic use: Culture cells without antibiotics or use 5- to 10-fold less antibiotics. Antibiotics bind serum proteins; the same concentration has a substantial impact on cells in serum-free media.
- Passaging: Use accutase or TrypLe in place of trypsin. If trypsin is used, inactivation must be completed by washing with media containing 1) FBS or 2) commercially available trypsin inhibitors. Extended incubation with trypsin or EDTA may impact adherence; monitor carefully.
- Filtering FRS™ Pioneer: FRS™ Pioneer is sterile and filtration is not required. Filtration risks disruption of lipids and cell cultures should be monitored.
- For maximum consistency: Prepare and store sterile basal media at 4°C. Warm basal media and add freshly-thawed FRS™ Pioneer directly prior to feed.
Optimising culture conditions:
- Use of a nutrient rich, complex basal medium is recommended. Examples include DMEM/F12 or RPMI.
- Monitor cell adherence carefully. Adherence challenges can manifest as morphology changes following a passage. To improve adherence, we recommend coating the plate with an appropriate adherence coating (see GECKO by Media City Scientific or recombinant vitronectin – VITRN0500).
- In some instances, conditioned media may improve results: Users may wish to trial retention of 10-25% conditioned media when feeding or passaging cells.
⚠️ Warning! Wear protective eyewear, gloves and clothing when conducting experiments
Instructions for storage and general use of FRS™ Pioneer
- FRS™ Pioneer should be diluted in basal cell culture media for culturing cells. More complex basal media may improve cell growth (e.g. DMEM/F12 in place of DMEM).
- FRS™ Pioneer is sterile and should be stored between -5°C and –20°C. Storage of FRS™ Pioneer in single use-aliquots is recommended to reduce freeze-thaw cycles.
- Do not allow FRS™ Pioneer to sit for extended periods of time in a water bath or incubator prior to use. Remove FRS Pioneer from the water bath immediately following thaw and warm only the media you intend to feed cells with.
- Cell culture should take place in a humidified atmosphere of 5% CO2.
- Cells may be frozen in freeze media made of 90% FRS™ Pioneer + 10% DMSO with results typically equivalent to 90% FBS + 10% DMSO.
- FRS™ Pioneer is for research or further manufacturing use only. If you seek to use FRS™ Pioneer in a clinical setting, please contact Media City Scientific.
Preparing FRS Pioneer
1. Thaw FRS™ Pioneer in the waterbath. Swirl regularly. Remove from heat immediately upon thaw.
2. Gently mix the thawed FRS™ Pioneer by inverting the container 2 to 3 times.
3. Prepare your basal media by sterile filtration if necessary.
4. Add sterile FRS™ Pioneer directly to your preferred basal media.
5. To maximise the consistency of your results, we recommend thawing the appropriate amount of FRS™ Pioneer and adding it to warmed basal media directly before feeding cells. FRS™ Pioneer is sterile and can be added directly to cell culture without filtration.
6. Alternative: Complete FRS™ Pioneer-containing media can be stored in the fridge for 2 weeks. When using FRS™ Pioneer-containing medium, warm only the aliquot you intend to use for feeding.
2. Gently mix the thawed FRS™ Pioneer by inverting the container 2 to 3 times.
3. Prepare your basal media by sterile filtration if necessary.
4. Add sterile FRS™ Pioneer directly to your preferred basal media.
5. To maximise the consistency of your results, we recommend thawing the appropriate amount of FRS™ Pioneer and adding it to warmed basal media directly before feeding cells. FRS™ Pioneer is sterile and can be added directly to cell culture without filtration.
6. Alternative: Complete FRS™ Pioneer-containing media can be stored in the fridge for 2 weeks. When using FRS™ Pioneer-containing medium, warm only the aliquot you intend to use for feeding.
Adaptation from FBS-Containing Media
Cells can be adapted directly or sequentially (weaned) to FRS™ Pioneer.
Direct adaptation
This approach is recommended for your first adaptation attempt, as it is successful for many cell types and cell lines. Thaw or passage cells in the usual FBS-containing media, then perform a 100% feed to FRS™ Pioneer-containing media after 24 hours when the cells are growing exponentially. White paper available for more details.
Sequential adaptation (weaning)
If direct adaptation does not support cell growth, we recommend a sequential adaptation. This weaning process gradually replaces serum containing media with FRS™ Pioneer-containing media. Whilst there is no prescribed speed for weaning, we recommend the media containing serum is halved at each passage until eliminated. For example:
Direct adaptation
This approach is recommended for your first adaptation attempt, as it is successful for many cell types and cell lines. Thaw or passage cells in the usual FBS-containing media, then perform a 100% feed to FRS™ Pioneer-containing media after 24 hours when the cells are growing exponentially. White paper available for more details.
Sequential adaptation (weaning)
If direct adaptation does not support cell growth, we recommend a sequential adaptation. This weaning process gradually replaces serum containing media with FRS™ Pioneer-containing media. Whilst there is no prescribed speed for weaning, we recommend the media containing serum is halved at each passage until eliminated. For example:
Users may choose to accelerate this transition for cells which are performing well or repeat a step if cell growth is diminished. Ultimately, the adaptation protocol adopted for cells should be reflective of a cell type’s particular requirements.
